摘要
根据鸭瘟病毒UL6和UL7基因序列,设计合成了一对引物,以2株疫苗株1、株强毒株和1株山东分离株DNA为模板,进行PCR扩增,得到预期690 bp的目的片段。将扩增的目的片段克隆到pMD18_T载体,经Amp平板筛选,HindIII、BamHI双酶切鉴定,获得阳性重组质粒。对重组质粒进行序列测定,与参考序列比较,山东分离株与参考序列的同源性为99.7%,其余3株DPV与参考序列的同源性均为100%。应用PCR可检测人工感染和自然感染鸭瘟的组织中的鸭瘟病毒,表明PCR检测鸭瘟病毒具有很高的特异性、敏感性,该法能够用于鸭瘟急性及亚临床感染的检测与诊断。
According to the UL6 and UL7 gene data ot duck plague viris（DPV） ,a pair ot prmers were designed and used for a polymerase chain reaction（PCR） with two duck plague vaccine strains and one standard strain and one field isolate. A specific 690bp DNA fragment was amplified,which was cloned into PMD18-T vector,and the positive ricombinant clone was selected by the Amp agar plate and characterized by Hind Ⅲ and BamH Ⅰ enzyme digestion. The recombinant plasmids were sequenced and compared with pulished sequence.The homology of Shandong isolated strain and the others was 99.7 % and 100 % respectively.The DPV specific DNA fragment was amplified from the infected organs samples of DPV. The PCR is specific and sensitive method which can be used for detection and diagnosis of latent and subclinical DPV infection.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2005年第5期408-411,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
山东省科技攻关项目(编号:012020104)
